JPH02243624A - Antiviral agent - Google Patents

Abstract

PURPOSE:To obtain an antiviral agent useful for treating virus infectious diseases, having excellent antiviral action and high safety, comprising a p- guanidinobenzoic acid derivative or a salt thereof as an active ingredient. CONSTITUTION:0.1-99%, preferably 0.5-90% based on total composition of a p-guanidinobenzoic acid derivative shown by formula I (R is group shown by formula II or formula III) or a salt thereof as an active ingredient is blended with a conventional drug carrier or additive and pharmaceutically manufactured by a conventional procedure to give the aimed substance. The substance may be prepared into forms for oral and parenteral administration such as tablet, troche, powder, pill, granule, capsule, suspension, syrup, injection, aerosol, oint ment or eye drop. The antiviral agent prevents plaque formation and viral multiplication in cavity solution of chorio-allantoic membrane of ovum gallinaceum especially on envelope virus such as human influenza virus.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is useful in the field of medicine. More specifically, the present invention relates to therapeutic agents for diseases caused by viral infections.

General formula % formula % (1) Formula: -C>CH, COOCH, CONCCH. ),
The compound represented by [representing a ler group] is a known substance and is used to treat acute pancreatitis such as acute pancreatitis, acute exacerbation of chronic pancreatitis, postoperative acute pancreatitis, acute W inflammation after pancreatography, and traumatic pancreatitis. Used in medical settings as therapeutic agents for symptoms [Japanese Patent Publication No. 54-41582, Japanese Patent Publication No. 54-41583, Japanese Patent Publication No. 61-1063, Pharmaceutical Research, No. 1
3, p. 756 (1982), BiocMm.Bioph
ys. Acta. ), Volume 484, Page 417 (19
1977), Japanese Journal of Pharmacology CJapan. J.

PharIIlacol. ), Vol. 41, p. 155 (1986) and Japanese Pharmacological Journal, Vol. 84, No. 36
See page 3 (1984)]. However, the general formula (1
) There is no report yet that the p-guanidinobenzoic acid derivative represented by the following formula shows antiviral activity.

A number of nucleic acid derivatives are known as antiviral agents, and amantadine and other anti-influenza virus agents are known. Antiviral drugs based on nucleic acid derivatives have side effects such as liver dysfunction, mutagenicity, and subacute toxicity, and amantadine has unfavorable properties such as side effects such as teratogenicity and decreased efficacy due to increased frequency of use. have [
Virology, published by Raven Press, pp. 323~
See page 348 (1985)]. It is desired to solve these problems of known antiviral agents.

As a result of intensive research aimed at solving the above-mentioned problems, the present inventors have found that p-guanidinobenzoic acid derivatives or non-toxic salts thereof represented by the general formula H) shown below are excellent antivirals. The present invention was completed after discovering that the method is superior in terms of effectiveness and safety.

That is, the present invention uses the general formula % formula % () Formula: OCH,COOCH,CON(CH,),
The present invention relates to an antiviral agent containing a p-guanidinobenzoic acid derivative or a nontoxic salt thereof as an active ingredient.

Examples of viral infections to which the antiviral agent of the present invention can be applied include infections caused by enveloped viruses such as influenza virus, rose influenza virus, herpes virus, and retroviruses such as human immunodeficiency virus (HIV). .

The non-toxic salt of the p-guanidinobenzoic acid derivative represented by the general formula (1) means a salt that can be used as a pharmaceutical, such as a salt with an inorganic acid such as hydrochloric acid, sulfuric acid or phosphoric acid, or a salt with an inorganic acid such as acetic acid, Salts with organic acids such as lactic acid, citric acid, methanesulfonic acid, p-toluenesulfonic acid, succinic acid, fumaric acid or maleic acid are mentioned.

The compound represented by the general formula (1) according to the present invention can be prepared by a known method, for example, as described in Japanese Patent Publication No. 54-41, 582, Japanese Patent Publication No. 54-41.583, Japanese Patent Publication No. 61-1063, etc. It can be manufactured and obtained by the following method.

Below, p-guanidine expressed by Ikki formula (1),
In order to concretely demonstrate the antiviral effect of aromatic acid derivatives, a test example will be given and explained.

(Left below) Test drug substance A: 6-amidino-2-naphthyl p-guanidinobenzoate nimethane sulfonate Substance B: N,N-dimethylcarbamoylmethyl p
-(p-guanidinobenzoyloxy)phenylacetate methanesulfonate Test Example 1 Inhibitory effect on black formation of human influenza virus Virology, Vol. 29,
The inhibitory effect of substance A and substance B of the present invention on the black formation of human influenza virus A/WSN was measured by the method described on pages 84 to 91 (1966).

In other words, IXIO' MDCK cells (human kidney cells) are
- plated in 6aTl plastic dishes containing MEM medium;
After culturing in a carbon dioxide incubator at 37°C for 24 hours, discard the medium and collect around 50 human influenza viruses A/1Js.
Added 0.1 m of virus dilution solution containing N (+). Shake well and leave at room temperature for 30 minutes. Add agar overlay solution containing a specified amount of test drug. After the agar solidifies, heat at 37°C. Cultured in a carbon dioxide incubator for 3 days. After culture, the agar layer was removed.
The cells were stained with a staining solution and the black number was counted. Amantadine was used as a control drug.

The black formation inhibition rate was calculated using the following formula, and the results are shown in Table 1.

T: Average black number in the presence of test drug C: Control group average black number [D-HEM medium] 1.9% Dulbecco's Modified E
AGLE MEDIUM “N15sui” powder 9.5g
(Hereinafter, one bottle of DMEM will be referred to as DMEM) and sterilized in an autoclave. This was sterilized by mouth, 0.584 g of glutamine, and 0.1 g of kanamycin.
g, 20 mfi of 7.5% aqueous sodium bicarbonate solution and 110 rnQ of immobilized fetal bovine serum.

[Dilution solution for viruses]

Phosphate Buffered 5aline
1 n of calcium chloride 10g, magnesium chloride 1
A solution containing 0g and 30% bovine serum albumin 5.8mQ.

[Agar multilayer solution]

Pure water 16m
Q2 wood DMEM +BA
50m A1% DEAE dextran 1
mf17.5% sodium hydrogen carbonate aqueous solution 2mQ2
．． 0% agar (noble) 30
m Q (2* DMEM 10 BA) 4 Dulbecco 2
50m QIM HEPES
10m f110% bovine serum 2
0mΩ7.5% sodium bicarbonate aqueous solution 16mΩ6
% glutamine aqueous solution 10m Q kanamycin aqueous solution (100Ing/m n ) 1m Q pure
Water 193mQ
[Staining solution] Dissolve 100 μm of crystal violet in 20111Ω ethanol and add 80 mΩ of water.

Table 1 As is clear from Table 1, substance A according to the present invention has a 5.0
Black formation of human influenza virus was inhibited by 52.1% at a concentration of μg 1mQ. Further, substance B according to the present invention inhibited the formation of a human influenza virus by 51.5% at a concentration of 20 μg/mQ.

Further, substance A and substance B according to the present invention each have 10
Black formation of human influenza virus was 100% inhibited at concentrations of μg1mQ, 100μglIIlΩ or higher. Amantadine, the control drug, was 30 μg/mΩ.
The black formation was inhibited by 60.1% at a concentration of . Therefore, substance A and substance B according to the present invention have a stronger inhibitory effect on the growth of human influenza virus than amantadine.

Test Example 2 Human influenza virus (A/Fukuoka/8
Inhibitory effect on the proliferation of 0) in the allantoic fluid of hatched chicken eggs on the 11th day. 0.1 ml of virus solution containing 6 x 10" Fukuoka virus was inoculated into the allantoic fluid of hatched chicken eggs. inoculate the drug), 37℃ for 2 days
Cultivate in an incubator. After culturing, the fetuses were killed by allowing them to stand overnight at 4°C, and the allantoic fluid was collected, and the HA titer of the virus was measured using chicken red blood cells. The results are shown in Table 2. The control group showed an HA titer of 532 HA (5X 10'PFA/if).

Table 2 As is clear from Table 2, Substance A and Substance B according to the present invention inhibit the proliferation of human influenza virus in the allantoic fluid of hatched chicken eggs more strongly than amantadine cited as a comparative example. I understand that.

Further, substance A and substance B according to the present invention were added at 50 μg/each to 3-day cultured cells of 1MDCK (Daiken Cells).
mQ showed no cytotoxicity up to a concentration of 1000 μg/rnQ, but amantadine clearly showed cytotoxicity, albeit slightly, even at 30 μg/mQ.

Therefore, substance A and substance B according to the present invention are compounds with a wider safety margin than amantadine.

When the compound of the present invention represented by the general formula N) is used as an antiviral agent, the dosage is usually 1 to 500 μg/dose, 1 to 3 times a day, rectally. For internal administration, 1 to 100 mg is administered 1 to 3 times a day, for inhalation, 0.1 to 500 mg is administered to the bronchi 2 to 3 times a day, for intravenous injection, Administer 0.1-10μ once or twice a day, or 0.1-50μ once a day for intranasal administration.
0■ administered 2 to 4 times, as eye drops 0.1 to 50 per dose
mg 3 to 4 times a day, 1 to 50 mg per dose as an ointment
It is recommended that the dose be administered 1 to 3 times, but doses outside these ranges may be used depending on age, sex, body weight, symptoms, etc.

The compound according to the present invention represented by general formula (I) can be made into a dosage form for oral or parenteral administration using the following formulation additives.

Unit dosage forms that can be used for oral administration include, for example, tablets, troches, powders, pills, granules, and capsules. These dosage forms include binders such as, for example, gum arabic, gelatin, sorbitol, tragacanth, polyvinylpyrrolidone, polyvinyl alcohol, hydroxypropylmethylcellulose, methylcellulose, crystalline cellulose or sodium carboxymethylcellulose, such as lactose, sugar, sucrose, sucrose, Excipients such as mannitol, corn starch, calcium phosphate, sorbitol or microcrystalline cellulose, lubricants such as magnesium stearate, talc, polyethylene glycol or silica, or calcium or carboxymethyl cellulose such as potato starch, low substituted hydroxypropylcellulose, carboxymethylcellulose, etc. Additives such as disintegrants such as sodium methyl starch may be used alone or in combination as appropriate. In addition, in soft capsules, commonly used vehicles such as vegetable oil, polyethylene glycol or glycerin, and the following oil-based suspensions may be used. The composition may contain a wetting agent such as a liquid agent, a liquid agent, or a surfactant.

Liquid formulations include, for example, aqueous or oily suspensions, solutions, syrups, elixirs, or lyophilizates that can be redissolved in water or other suitable vehicle at the time of use. The liquid preparations include suspending agents such as methylcellulose, sodium carboxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, tragacanth, gelatin or sodium alginate;
For example, lecithin, sorbitan, fatty acid esters, emulsifiers such as gum arabic or tragacanth, such as polyoxyethylene sorbitan fatty acid ester, polyoxyethylene fatty acid ester, hydrogenated castor oil, sesame oil, soybean oil, propylene glycol, polyethylene glycol or ethyl alcohol. humectants, preservatives such as methyl p-hydroxybenzoate, propyl p-hydroxybenzoate or sorbic acid, or additives such as sweeteners such as simple syrup, sucrose, sorbitol or mannitol, alone or in appropriate combinations. You may also use it.

The base for rectal administration may be an oily base such as cacao butter, lytepsol or triglyceride, or a water-soluble base such as glycerin, roselatin or macrogol.

Examples of additives for injections include polyoxyethylene,
solubilizing agents such as hydrogenated castor oil or sodium benzoate; tonicity agents such as dextrose, sodium chloride or glycerin; or stabilizers such as sodium sulfite, anhydrous sodium sulfite, sodium metabisulfite or glycerin. These may be used alone or in appropriate combination.

When administering to the respiratory tract such as the nose and bronchi, dosage forms such as aerosols, inhalation solutions, liquids, powders, capsules, or ointments can be used. In the case of aerosols, non-ionic surfactants such as Aracel or Span 80, amphoteric surfactants such as lecithin or dispersants such as oleyl alcohol and for example butane or Freon 0 are used.
or an aqueous aerosol formulation consisting of an isotonic agent such as physiological saline, phosphate buffer or acetate buffer, and purified water or distilled water for injection. can.

In the case of liquid preparations, for example, polyethylene glycol, sorbitol, polysorbate, physiological saline, or the like can be used as a pharmaceutical carrier. In the case of a powder, for example, crystalline cellulose, α-cellulose, cross-linked sodium carboxymethyl cellulose, hydroxypropyl cellulose, carboxymethyl starch or amylose can be used as a pharmaceutical carrier. Furthermore, this powder can be filled into capsules to prepare capsules. In the case of ointments, for example, polyethylene glycol, hydroxyethylcellulose, methylcellulose, carboxymethylcellulose or hydroxypropylcellulose can be used as the pharmaceutical carrier. Furthermore, it is also possible to form a dosage form that adheres to the mucous membranes of the oral cavity or nose and gradually releases the active ingredient. Examples of bases used in this dosage form include methylcellulose, ethylcellulose, propylcellulose, cellulose ethers such as droxyethylcellulose, carboxyethylcellulose, or hydroxypropylcellulose, or polyacrylic acid or carboxyvinyl polymers. It will be done.

These formulations can be prepared by respective known manufacturing methods.

The p-guanidinobenzoic acid derivatives of the invention are present in these formulations in an amount of about 0.1 to 99%, preferably 0.5 to 99% of the total composition.
It is preferable to contain 0%.

EXAMPLES The present invention will be specifically described below with reference to Examples, but the present invention is not limited thereto.

Example 1 6-amidino-2-naphthyl p-guanidinobenzoate 10 g of nimethane sulfonate, 400,300 g of polyethylene glycol and 800.5 g of polysorbate)
The mixture was heated and stirred to uniformly dissolve the mixture to produce a nasal solution. This preparation is administered intranasally in a dose of 0.1 ml.

Example 2 2 g of 6-amidino-2-naphthyl p-guanidinobenzoate nimethane sulfonate was dissolved in physiological saline, and the whole was dissolved in 1. A liquid preparation was prepared as OOOmQ. This preparation is placed in a nasal dropper and administered as a spray at a dose of approximately 0.2 mΩ each time.

Example 3 2 g of 6-amidino-2-naphthyl p-guanidinobenzoate nimethane sulfonate is dissolved in 500 ml of distilled water for injection heated to 50°C. Add 400 mfl of 12.5% sorbitol aqueous solution to this, mix uniformly, and adjust the pH to 4.0 with 0.1N hydrochloric acid. After that, add distilled water for injection and mix the whole with 1. A liquid preparation was prepared as OOOmQ. This solution is placed in a nasal spray container and administered as a spray at a dose of approximately 0.2 mQ at a time.

Example 4 After pulverizing 1.0 g of 6-amidino-2-naphthyl p-guanidinobenzoate nimethane sulfonate in an agate mortar, 85,200 mg of Span was added and kneaded uniformly. After cooling the resulting mixture to -40°C, it was mixed with 10 g of a 4:l mixture of Freon 11, Freon 12 and Freon 114 previously cooled to -40°C, and then cold-filled into a pressure-resistant container to form an aerosol. Obtain the formulation.

Example 5 50 mg of 6-amidino-2-naphthyl p-guanidinobenzoate nimethane sulfonate and 450 mg of crystalline cellulose are thoroughly ground in a mortar to obtain a uniform powder composition. A capsule preparation for nasal administration was obtained by filling this powder composition into a predetermined capsule.

Example 6 10 μL of freeze-dried powder of rhoamidino 2-naphthyl p-guanidinobenzoate nimethane sulfonate was added to 5
An intravenous injection solution was obtained by dissolving in 500mM % glucose injection solution. This injection solution can be instilled intravenously over a period of about 2 hours once or twice a day.

Example 7 Rhoamidino 2-naphthyl p-guanidinobenzoate nimethane sulfonate 10 g, white sugar (fine powder) 1
00g and 7g of gum arabic (fine powder) were mixed, and an appropriate amount of purified water was added and kneaded. After drying this kneaded product, 1 g of magnesium stearate was added and compression molded by a conventional method to obtain 100 troches.

Example 8 N,N-dimethylcarbamoylmethyl p-(p-guanidinobenzoyloxy)phenylacetate methanesulfonate 20g, polyethylene glycol 40020
0 g and 800.5 g of polysorbate were uniformly dissolved with heating and stirring to produce a nasal solution. This preparation is administered by dropping 0.1 mQ into the nasal cavity once.

Example 9 6-amidino-2-naphthyl p-guanidinobenzoate 50 mg of nimethane sulfonic acid and 450 mg of hydroxypropyl cellulose were thoroughly ground in a mortar,
A homogeneous powder composition is obtained. A capsule preparation for nasal administration was obtained by filling this powder composition into a predetermined capsule.

Example 1O 200 n+g of N,N-dimethylcarbamoylmethyl p-(p-guanidinobenzoyloxy)phenylacetate methanesulfonate and 100 mg of hydroxypropyl cellulose are thoroughly ground in a mortar to obtain a uniform powder composition. A capsule preparation for nasal administration was obtained by filling this powder composition into a predetermined capsule.

Example 11 6-amidino-2-naphthyl p-guanidinobenzoate 500 g of nimethane sulfonic acid and 1 g of hydroxypropylcellulose 50II were dissolved in 5 mQ of distilled water for injection, and then freeze-dried to obtain a uniform composition. A capsule preparation for nasal administration was obtained by filling this composition into a predetermined capsule.

Example 12 50 mg of N,N-dimethylcarbamoylmethyl p-(p-guanidinobenzoyloxy)phenylacetate methanesulfonate and 50+ ng of hydroxypropyl cellulose were dissolved in 5 mfl of distilled water for injection, and then lyophilized to obtain a uniform composition. Ta. A capsule preparation for nasal administration was obtained by filling this composition into a predetermined capsule.

Example 13 6-amidino-2-naphthyl p-guanidinobenzoate 10 g of nimethane sulfonic acid, 25 g of hydroxypropyl cellulose, and 93,420 g of Carbobol were mixed uniformly, and 0.0 g of magnesium stearate was added to the mixture.
5 g was added and compression molded by a conventional method to obtain 100 flat tablets. This preparation is used by attaching one piece to the oral mucosa at a time.

Since the substance of the present invention inhibits the black formation of enveloped viruses such as human influenza virus and the proliferation of the virus in the chorioallantoic fluid of chicken eggs, the present invention is effective against viral infections. It contributes to treatment.

Claims (6)

[Claims] (1) General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (I) [In the formula, R is a formula: ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ Groups or formulas represented by: ▲ Numerical formulas, chemical formulas, tables, etc. An antiviral agent characterized by containing a p-guanidinobenzoic acid derivative represented by ▼ or a nontoxic salt thereof as an active ingredient. (2) Formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (A) A first product characterized by containing 6-amidino-2-naphthyl p-guanidinobenzoate or a non-toxic salt thereof as an active ingredient. The antiviral agent according to the claims. (3) Formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼ (B) N,N-dimethylcarbamoylmethyl p-(
The antiviral agent according to claim 1, which contains p-guanidinobenzoyloxy)phenylacetate or a nontoxic salt thereof as an active ingredient. (4) General formula for pharmaceutical carrier and effective amount ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (I) [In the formula, R is the formula: ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ Group or formula The antiviral agent according to claim 1, comprising a p-guanidinobenzoic acid derivative or a non-toxic salt thereof, represented by ▲ ▲ represents a group represented by ▼, which includes numerical formulas, chemical formulas, tables, etc.]. (5) The antiviral agent according to claim 4, wherein the pharmaceutical carrier is a pharmaceutical carrier for nasal administration. (6) General formula for effective amount ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ (I) [In the formula, R is a formula: ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ A group or formula represented by: ▲ Numerical formula, A method for treating viral infections, which comprises administering a p-guanidinobenzoic acid derivative or a nontoxic salt thereof, represented by the following chemical formula, table, etc.